Perforated patch clamp recording was used to study the control of membrane potential (V(m)) and spontaneous electrical activity in the rat pinealocyte by norepinephrine. Norepinephrine did not alter spiking frequency. However, it was found to act through α(1B)-adrenoreceptors in a concentration-dependent manner (0.1-10 μM) to produce a biphasic change in V(m). The initial response was a hyperpolarization (∼13 mV from a resting potential of -46 mV) due to a transient (∼5 sec) outward K(+) current (∼50 pA). This current appears to be triggered by Ca(2+) released from intracellular stores, based on the observation that it was also seen in cells bathed in Ca(2+)-deficient medium. In addition, pharmacological studies indicate that this current was dependent on phospholipase C (PLC) activation and was in part mediated by bicuculline methiodide and apamin-sensitive Ca(2+)-controlled K(+) channels. The initial transient hyperpolarization was followed by a sustained depolarization (∼4 mV) due to an inward current (∼10 pA). This response was dependent on PLC-dependent activation of Na(+)/Ca(2+) influx but did not involve nifedipine-sensitive voltage-gated Ca(2+) channels. Together, these results indicate for the first time that activation of α(1B)-adrenoreceptors initiates a PLC-dependent biphasic change in pinealocyte V(m) characterized by an initial transient hyperpolarization mediated by a mixture of Ca(2+)-activated K(+) channels followed by a sustained depolarization mediated by a Ca(2+)-conducting nonselective cation channel. These observations indicate that both continuous elevation of intracellular Ca(2+) and sustained depolarization at approximately -40 mV are associated with and are likely to be required for activation of the pinealocyte.

• MeSH
• Receptors, Adrenergic, alpha-1 physiology   MeSH
• Pineal Gland cytology   drug effects   physiology   MeSH
• Type C Phospholipases physiology   MeSH
• Rats MeSH
• Membrane Potentials drug effects   MeSH
• Norepinephrine pharmacology   MeSH
• Rats, Sprague-Dawley MeSH
• Calcium metabolism   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Intramural MeSH

Non-dioxin-like polychlorinated biphenyls (NDL-PCBs) have been shown to act as tumor promoters in liver; however, the exact mechanisms of their action are still only partially understood. One of the interesting effects of NDL-PCBs is the acute inhibition of gap junctional intercellular communication (GJIC), an effect, which has been often found to be associated with tumor promotion. As previous studies have suggested that NDL-PCB-induced disruption of lipid signalling pathways might correspond with GJIC inhibition, we investigated effects of PCBs on the release of arachidonic acid (AA) in the rat liver epithelial WB-F344 cell line, a well-established model of liver progenitor cells. We found that both 2,2',4,4'-tetrachlorobiphenyl (PCB 47) and 2,2',4,4',5,5'-hexachlorobiphenyl (PCB 153), but not the dioxin-like, non-ortho-substituted, 3,3',4,4',5-pentachlorobiphenyl (PCB 126), induce a massive release of AA. The AA release, induced by PCB 153, was partially inhibited by extracellular signal-regulated kinases 1/2 (ERK1/2) signalling inhibitor, U0126, and by cytosolic phospholipase A(2) (cPLA(2)) inhibitor, AACOCF(3). Although PCB 153 induced both ERK1/2 and p38 activation, the specific p38 kinase inhibitor, SB203580, had no effect on AA release. Inhibitors of other phospholipases, including phosphatidylcholine-specific phospholipase C or phosphatidylinositol-specific phospholipase C, were also without effect. Taken together, our findings suggest that the AA release, induced by non-dioxin-like PCBs in liver progenitor cell line, is partially mediated by cytosolic PLA(2) and regulated by ERK1/2 kinases. Our results suggest that more attention should be paid to cell signalling pathways regulated by AA or eicosanoids after PCB exposure, which might be involved in their toxic effects.

• MeSH
• Cell Line MeSH
• Phospholipases A2, Cytosolic metabolism   drug effects   MeSH
• Epithelial Cells metabolism   drug effects   MeSH
• Financing, Organized MeSH
• Liver cytology   metabolism   drug effects   MeSH
• Stem Cells metabolism   drug effects   MeSH
• Rats MeSH
• Arachidonic Acids metabolism   MeSH
• Environmental Pollutants toxicity   MeSH
• Mitogen-Activated Protein Kinase 1 metabolism   drug effects   MeSH
• Mitogen-Activated Protein Kinase 3 metabolism   drug effects   MeSH
• Polychlorinated Biphenyls pharmacology   toxicity   MeSH
• Rats, Inbred F344 MeSH
• Signal Transduction drug effects   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH

CD148 is a receptor-like protein-tyrosine phosphatase known to inhibit transduction of mitogenic signals in non-hematopoietic cells. Similarly, in the hematopoietic lineage, CD148 inhibited signal transduction downstream of T cell receptor. However, it also augmented immunoreceptor signaling in B cells and macrophages via dephosphorylating C-terminal tyrosine of Src family kinases (SFK). Accordingly, endogenous CD148 compensated for the loss of the main SFK activator CD45 in murine B cells and macrophages but not in T cells. Hypothetical explanations for the difference between T cells and other leukocyte lineages include the inability of CD148 to dephosphorylate a specific set of SFKs involved in T cell activation or the lack of CD148 expression during critical stages of T cell development. Here we describe striking differences in CD148 expression between human and murine thymocyte subsets, the only unifying feature being the absence of CD148 during the positive selection when the major developmental block occurs under CD45 deficiency. Moreover, we demonstrate that similar to CD45, CD148 has both activating and inhibitory effects on the SFKs involved in TCR signaling. However, in the absence of CD45, activating effects prevail, resulting in functional complementation of CD45 deficiency in human T cell lines. Importantly, this is independent of the tyrosines in the CD148 C-terminal tail, contradicting the recently proposed phosphotyrosine displacement model as a mechanism of SFK activation by CD148. Collectively, our data suggest that differential effects of CD148 in T cells and other leukocyte subsets cannot be explained by the CD148 inability to activate T cell SFKs but rather by its dual inhibitory/activatory function and specific expression pattern.

• MeSH
• Adaptor Proteins, Signal Transducing metabolism   MeSH
• Enzyme Activation MeSH
• Leukocyte Common Antigens deficiency   MeSH
• Phospholipase C gamma metabolism   MeSH
• Phosphorylation MeSH
• Hematopoietic Stem Cells cytology   enzymology   metabolism   MeSH
• Jurkat Cells MeSH
• Humans MeSH
• Membrane Proteins metabolism   MeSH
• Mice MeSH
• Receptors, Antigen, T-Cell immunology   metabolism   MeSH
• Gene Expression Regulation, Enzymologic MeSH
• Signal Transduction MeSH
• src-Family Kinases chemistry   metabolism   MeSH
• T-Lymphocytes cytology   enzymology   metabolism   MeSH
• Protein Structure, Tertiary MeSH
• Thymus Gland cytology   MeSH
• Tyrosine metabolism   MeSH
• Receptor-Like Protein Tyrosine Phosphatases, Class 3 chemistry   genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH

Long-term potentiation (LTP) in the rat hippocampus is the most extensively studied cellular model for learning and memory. Induction of classical LTP involves an NMDA-receptor- and calcium-dependent increase in functional synaptic AMPA receptors, mediated by enhanced recycling of internalized AMPA receptors back to the postsynaptic membrane. Here we report a physiologically relevant NMDA-receptor-independent mechanism that drives increased AMPA receptor recycling and LTP. This pathway requires the metabotropic action of kainate receptors and activation of G protein, protein kinase C and phospholipase C. Like classical LTP, kainate-receptor-dependent LTP recruits recycling endosomes to spines, enhances synaptic recycling of AMPA receptors to increase their surface expression and elicits structural changes in spines, including increased growth and maturation. These data reveal a new and, to our knowledge, previously unsuspected role for postsynaptic kainate receptors in the induction of functional and structural plasticity in the hippocampus.

• MeSH
• Receptors, AMPA metabolism   MeSH
• Dendritic Spines metabolism   MeSH
• Long-Term Potentiation physiology   MeSH
• Endosomes metabolism   MeSH
• Type C Phospholipases metabolism   MeSH
• Hippocampus physiology   MeSH
• Rats MeSH
• Cells, Cultured MeSH
• Neurons metabolism   physiology   MeSH
• Protein Kinase C metabolism   MeSH
• GTP-Binding Proteins metabolism   MeSH
• Receptors, Kainic Acid physiology   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Mammal heart tissue has long been assumed to be the exclusive domain of the M(2) subtype of muscarinic receptor, but data supporting the presence of other subtypes also exist. We have tested the hypothesis that muscarinic receptors other than the M(2) subtype are present in the heart as minor populations. We used several approaches: a set of competition binding experiments with pirenzepine, AFDX-116, 4-DAMP, PD 102807, p-F-HHSiD, AQ-RA 741, DAU 5884, methoctramine and tripinamide, blockage of M(1) muscarinic receptors using MT7 toxin, subtype-specific immunoprecipitation experiments and determination of phospholipase C activity. We also attempted to block M(1)-M(4) receptors using co-treatment with MT7 and AQ-RA 741. Our results show that only the M(2) subtype is present in the atria. In the ventricles, however, we were able to determine that 20% (on average) of the muscarinic receptors were subtypes other than M(2), with the majority of these belonging to the M(1) subtype. We were also able to detect a marginal fraction (6 +/- 2%) of receptors that, based on other findings, belong mainly to the M(5) muscarinic receptors. Co-treatment with MT7 and AQ-RA 741 was not a suitable tool for blocking of M(1)-M(4) receptors and can not therefore be used as a method for M(5) muscarinic receptor detection in substitution to crude venom. These results provide further evidence of the expression of the M(1) muscarinic receptor subtype in the rat heart and also show that the heart contains at least one other, albeit minor, muscarinic receptor population, which most likely belongs to the M(5) muscarinic receptors but not to that of the M(3) receptors.

• MeSH
• Gene Expression MeSH
• Financing, Organized MeSH
• Type C Phospholipases metabolism   MeSH
• Binding, Competitive MeSH
• Rats MeSH
• Rats, Wistar MeSH
• Receptor, Muscarinic M1 metabolism   drug effects   MeSH
• Receptor, Muscarinic M2 metabolism   drug effects   MeSH
• Receptor, Muscarinic M3 metabolism   drug effects   MeSH
• Receptor, Muscarinic M5 metabolism   drug effects   MeSH
• Receptors, Muscarinic metabolism   drug effects   MeSH
• Heart Ventricles metabolism   drug effects   MeSH
• Heart Atria metabolism   drug effects   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH

Lupénka je systémové zánětlivé onemocnění. V patogenezi se uplatňují genetické faktory a vlivy vnějšího prostředí. Poslední studie ukazují na souvislost lupénky s metabolickým syndromem a kardiovaskulárními onemocněními. V naší práci jsme zkoumali změny sérových hladin vybraných ukazatelů zánětu, jako jsou: C-reaktivní protein (CRP), fosfolipáza A2 asociovaná s lipoproteiny (Lp-PLA2), adiponektin, leptin, lipokalin 2, rezistin a retinol vázající protein 4 (RBP4). Sérové ukazatele byly měřeny a porovnávány u 32 pacientů se středně závažnou až závažnou formou lupénky (medián PASI 17,1) a u 24 zdravých kontrol (dárci krve). U pacientů s lupénkou byly zjištěny signifikantně vyšší hladiny CRP, Lp-PLA2, lipokalinu 2 a leptinu v porovnání s kontrolní skupinou. U pacientů s lupénkou byla prokázána významná negativní korelace mezi body mass indexem (BMI) a adiponektinem, pozitivní korelace byly zjištěny mezi BMI a leptinem a mezi BMI a lipokalinem 2. Ukazatelé CRP, Lp-PLA2, lipokalin 2 a leptin můžou sloužit k posouzení přítomnosti systémového postižení při lupénce a k včasnému zachycení a léčení komorbidit lupénky.

Psoriasis is a systemic inflammatory disease. Genetic and environmental factors play a role in its pathogenesis. Recent studies refer to association between psoriasis with metabolic syndrome and cardiovascular disease. In our study we examined changes in serum levels of selected indicators of inflammation such as C-reactive protein (CRP), lipoprotein associated phospholipase A2 (LpPLA2), adiponectin, leptin, lipocalin 2, resistin and retinol binding protein 4 (RBP4). Serum indicators were measured and compared in 32 patients with moderate to severe psoriasis (median PASI 17,1) and in 24 healthy controls (donors of blood). Significantly higher levels of CRP, LpPLA2, lipocalin 2 and leptin were found in patients with psoriasis compared to the control group. Negative correlation were observed between body mass index BMI and adiponectin, positive correlations were found between BMI and leptin and between BMI and lipocalin 2 in patients with psoriasis. Indicators like CRP, LpPLA2, lipocalin 2 and leptin could be used to asses the systemic involvement in psoriasis and to recognition and treatment of psoriatic comorbidities early.

• Keywords
• fosfolipáza A2 asociovaná s lipoproteiny, retinol vázající protein 4, lipokalin 2,
• MeSH
• 1-Alkyl-2-acetylglycerophosphocholine Esterase blood   MeSH
• Adiponectin blood   MeSH
• Biomarkers * blood   MeSH
• C-Reactive Protein analysis   MeSH
• Body Mass Index MeSH
• Smoking MeSH
• Leptin blood   MeSH
• Middle Aged MeSH
• Humans MeSH
• Lipocalins blood   MeSH
• Psoriasis * blood   MeSH
• Resistin blood   MeSH
• Statistics as Topic MeSH
• Case-Control Studies MeSH
• Vitamin A blood   MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Female MeSH

Psoriáza je chronické recidivující zánětlivé onemocnění postihující 2–3 % evropské populace. Metabolický syndrom (MetS) je souborem centrální obezity, dyslipidémie, arteriální hypertenze a inzulinové rezistence a je rizikovým faktorem kardiovaskulárních chorob a diabetes mellitus II. typu (DM2T), jehož prevalence se pohybuje mezi 25–30 % v ekonomicky vyspělých zemích. Kouření je úzce spojeno s MetS: kuřáci mají zvýšené riziko MetS. Pacienti s lupénkou mají na straně jedné vyšší sklon ke kouření a na straně druhé také vyšší riziko rozvoje MetS. Jedním z mechanismů pojících psoriázu s MetS, může být subklinický zánět a zvýšená tvorba adipokinů, které jsou přítomny u obou nemocí. Kouření je spojeno s rozvojem oxidačního stresu a aktivací zánětlivých pochodů podporujících chronický průběh u psoriázy. Cílem studie bylo porovnat zánětlivé parametry: C-reaktivní protein (CRP), leptin, adiponektin, resistin a fosfolipázu A2 asociovanou s lipoproteiny (LpPLA-2) společně se složkami MetS u psoriatických pacientů (PP) a kontrolní skupiny (KS), s ohledem na přítomnost MetS a na kuřáckou anamnézu. Statisticky nevýznamně vyšší sklon ke kouření měla skupina PP, kde kouřilo 40,5 % v porovnání s KS, kde kouřilo jen 30,8 % jedinců. Dále jsme, u pacientů kuřáků bez MetS, v porovnání s pacienty nekuřáky bez MetS, shledali statisticky významně vyšší hodnoty PASI (Psoriasis Area and Severity Index, p < 0,5). Toto zjištění podporuje souvislost mezi závažností lupénky a kouřením bez ohledu na přítomnost MetS. Prozánětlivý ukazatel CRP (p < 0,01) a leptin (p < 0,05) a proaterogenní Lp-PLA2 (p <0,01) byly statisticky významně vyšší ve skupině nekuřáků PP s MetS, v porovnání s nekuřáky KS s MetS. Hladiny CRP (p = 0,001), LpPLA2 (p< 0,001), leptinu (p = 0,006) a resistinu (p = 0,005) byly statisticky významně ovlivněny lupénkou. Lupénka a obezita podle naší studie spolu souvisí pro statisticky významně vyšší BMI (p < 0,05) u nekuřáků PP s MetS v porovnání s nekuřáky KS s MetS. Závěrem můžeme potvrdit, že lupénka je zánětlivý stav, který jistě souvisí s obezitou bez ohledu na kuřáckou anamnézu. Kouření však zhoršuje závažnost lupénky.

Psoriasis is a chronic relapsing inflammatory disease affecting 2–3% of European population. The components of metabolic syndrome (MetS) include obesity, dylipidemia, high blood pressure and insulin resistence. MetS can increase the risk of cardiovascular disease and diabetes mellitus type II (DM2T) which prevalence is 25–30% in developed countries. Smoking is closely related to MetS: smokers are at higher risk of MetS. Patients with psoriasis are more likely to be smokers and have higher risk of developing MetS. Link between psoriasis and MetS can be explained by subclinical inflammation and elevation of adipokines levels, present in both diseases. Smoking activates inflammatory processes via oxidative stress that contributes to chronicity of psoriasis. The aim of this study was to compare inflammatory markers – C-reactive protein (CRP), leptin, adiponectin, resistin and lipoprotein associated phospholipase A2 (LpPLA2) as well as the factors of MetS between patients with psoriasis (PP) and the control group (CG) witht respect to positivity of MetS and smoking history. PP have a statistically non-significant higher tendency to smoke. In the PP group 40.5% of patients smoked and in the control group only 30.8% of individuals. Next, significantly higher psoriasis area and severity index (PASI, p < 0.05) was measured in patients without MetS, who smoked in comparison to non-smoking controls without MetS. This finding confirm a direct link between psoriasis and smoking regardless of the presence of MetS. We detected significantly higher levels of pro-inflammatory markers CRP (p < 0.01) and leptin (p < 0.05) and proatherogenic marker Lp-PLA2 (p < 0.01) in the group of PP non-smokers suffering with MetS in comparsion to non-smoking CG suffering with MetS. Levels of CRP (p = 0.001), Lp-PLA2 (p < 0.001), leptin (p = 0.006) and resistin (p = 0.005) were statistically significantly related to psoriasis. According to our study psoriasis and obesity are related due to significantly higher BMI (p < 0.05) in PP nonsmokers with MetS in comparsion to CG nonsmokers with MetS. In conclusion we can state that psoriasis is an inflammatory state related to obesity regardless of smoking history, but the severity of psoriasis is aggravated by smoking.

• MeSH
• 1-Alkyl-2-acetylglycerophosphocholine Esterase blood   MeSH
• Adiponectin blood   MeSH
• C-Reactive Protein analysis   MeSH
• Comorbidity MeSH
• Cigarette Smoking * adverse effects   MeSH
• Leptin blood   MeSH
• Humans MeSH
• Metabolic Syndrome diagnosis   MeSH
• Obesity MeSH
• Prevalence MeSH
• Psoriasis * diagnosis   MeSH
• Resistin blood   MeSH
• Risk Factors MeSH
• Check Tag
• Humans MeSH
• Publication type
• Clinical Trial MeSH
• Research Support, Non-U.S. Gov't MeSH

The multitalented neuropeptide galanin was first discovered 30 years ago but initially no biologic activity was found. Further research studies discovered the presence of galanin in the brain and some peripheral tissues, and galanin was identified as a modulator of neurotransmission in the central and peripheral nervous system. Over the last decade there were performed very intensive studies of the neuronal actions and also of nonneuronal actions of galanin. Other galanin family peptides have been described, namely galanin, galanin-like peptide, galanin-message associated peptide and alarin. The effect of these peptides is mediated through three galanin receptors subtypes, GalR1, GalR2 and GalR3 belonging to G protein coupled receptors, and signaling via multiple transduction pathways, including inhibition of cyclic AMP/protein kinase A (GalR1, GalR3) and stimulation of phospholipase C (GalR2). This also explains why one specific molecule of galanin can be responsible for different roles in different tissues. The present review summarizes the information currently available on the relationship between the galaninergic system and known pathological states. The research of novel galanin receptor specific agonists and antagonists is also very promising for its future role in pharmacological treatment. The galaninergic system is important target for current and future biomedical research.

• MeSH
• Cell Membrane metabolism   pathology   MeSH
• Galanin physiology   MeSH
• Humans MeSH
• Brain metabolism   pathology   MeSH
• Neurons metabolism   pathology   MeSH
• Receptors, Galanin physiology   MeSH
• Protein Structure, Secondary MeSH
• Signal Transduction physiology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

• MeSH
• Brain MeSH
• Neurophysiology MeSH
• Neurons MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• neurologie
• neurovědy

BACKGROUND: CD16 was previously suggested to be a new marker of basophils that is subject to downregulation by FcεRI crosslinking. Certain compounds, including supraoptimal concentrations of the PKC inhibitors, bisindolylmaleimides, decouple the release of granules containing CD203c, CD63 and histamine, and may thus help to identify the mechanisms related to the CD16 externalization. OBJECTIVE: We hypothesized that CD16 is differentially expressed on the surface of basophils in patients with birch pollen or insect venom allergy and is subject to a regulation in response to allergens. We also employed CD203c and CD63 externalization decoupling by bisindolylmaleimides. METHODS: We performed a basophil activation test coupled with CD16 and histamine detection using cells isolated from patients with allergy to birch pollen or insect venom and negative controls. We employed two PKC inhibitors, bisindolylmaleimide II and Ro 31-8220 at their supraoptimal concentrations and, after difficulties reproducing previously published data, we analyzed the fluorescence of these inhibitors alone. We identified the CD16 isoforms by sequencing nested RT-PCR amplicons from flow cytometry sorted basophils and by cleaving the CD16b GPI anchor using a phospholipase C. RESULTS: We provide the first evidence that CD16a is expressed as a surface antigen on a small subpopulation of human basophils in patients with respiratory and insect venom allergy, and this antigen shows increased surface expression following allergen challenge or FcεRI crosslinking. We rejected the apparent decoupling of the surface expression of basophil activation markers following the administration of bisindolylmaleimides. CONCLUSIONS & CLINICAL RELEVANCE: The inclusion of αCD16 in negative selection cocktails selects against a subset of basophils that are CD16+ or CD16dim . Using CD16dim basophils and unstained leucocytes, we show that previous studies with supraoptimal concentrations of bisindolylmaleimides are likely flawed and are not associated with the differential expression of CD203c and CD63.

• MeSH
• Hypersensitivity immunology   pathology   MeSH
• Tetraspanin 30 immunology   MeSH
• Basophils immunology   pathology   MeSH
• Adult MeSH
• Phosphoric Diester Hydrolases immunology   MeSH
• GPI-Linked Proteins immunology   MeSH
• Indoles chemistry   MeSH
• Arthropod Venoms toxicity   MeSH
• Insect Bites and Stings immunology   pathology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Maleimides chemistry   MeSH
• Pyrophosphatases immunology   MeSH
• Receptors, IgG immunology   MeSH
• Aged MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH